Histology Stains
Introduction
This page gives a general overview of some histological stains used to identify structures in cells and tissues. This stains information should also be considered in relation to Histology Fixatives. To see related histology images use the Category:Histology link.
Medicine Foundations students do not need to know stain information in this detail.
Common Stains and Their Reactions
Common Stains and Their Reactions | |||||
Haematoxylin | mucins - light blue | ||||
Eosin | colloid - pinkmuscle - red | ||||
Iron Haematoxylin | |||||
Van Gieson | muscle: yellow/browncartilage - pink | ||||
Verhoeff's Elastin | elastic fibres - black | ||||
Tartrazine | |||||
Silver Impregnation | reticular fibres - black | ||||
Methyl Green | |||||
Nuclear Fast Red | |||||
Gomori's Trichrome | keratin - redmuscle - purple/red | ||||
Heidenhain's Azan | muscle - red | ||||
Osmium Tetroxide | myelin, lipids - black | ||||
Alcian Blue | mucins, - blue | ||||
Periodic acid-Schiff (PAS) | mucins, glycogen, glycocalyx - magenta | ||||
Phosphotungstic Acid-Hematoxylin (PTAH) | muscle bands - blue | ||||
Masson's Trichrome | cartilage, mucins - blue or green; muscle - red | ||||
Luxol Fast Blue | myelin - blue | ||||
Aldehyde Fuchsin | elastic fibres, mast cells - deep purple | ||||
Light Green | |||||
Gallocyanin | nucleic acids, Nissl granules - dark blue | ||||
Romanowsky(e.g. Leishman's stain) | acidophils - redbasophils - blueazurophilic - purple | ||||
Aldehyde Pararosanilin | elastic fibres - purple |
Stain Templates |
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These templates put a link to this page and the appropriate stain text. Use the text in the curly brackets to insert the text in the curved brackets with a link to the Histology Stains page.
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Alcian Blue
- Stains mucopolysaccharides or glycosaminoglycans
- cationic dye (positively charged molecule) for the demonstration of glycosaminoglycans.
- binds anionic (negative) sites on the polysaccharide.
- can be combined with H&E and VG staining methods.
Alizarine Blue
Alizarine Brilliant Blue R
Alizarin Red
- Useful for identifying bone or other hight calcium structures.
- Stains insoluble calcium cations
- bright red stain
- Other metals such as barium, aluminium, mercury and magnesium (dark red)
Chromosome Banding
The term refers to the light and dark pattern, seen after staining with a dye, of individual chromosomes identified in metaphase. It is only in meiosis and mitosis during metaphase that chromosomes can be easily identified, during the normal cell life (interphase) the chromosomes are unravelled and distributed within the nucleus in chromosome territories. A band is that part of a chromosome which is clearly distinguishable from nearby regions by appearing darker or brighter with one or more banding techniques.
Depending on the type of stain used a number of different banding patterns can be seen:
- G-banding - banding pattern seen by treating with trypsin and then staining with the dye giemsa.
- R-banding - banding pattern seen as a of reverse giemsa chromosome banding, producing bands complementary to G-bands often used to determine whether there are deletions. Can be fluorescent using the dye acridine orange.
- Q-banding - banding pattern seen by treating with a fluorochrome or the fluorescent dye quinacrin.
- C-banding - banding pattern seen for centromeric or constitutive heterochromatin, the centromere appears as a stained band compared to other regions.
Metaphase is a cell division term referring to the third mitotic stage, mitotic spindle kinetochore microtubules align chromosomes in one midpoint plane. Metaphase ends when sister kinetochores separate. Originally based on light microscopy of living cells and electron microscopy of fixed and stained cells. A light microscope analysis called a "metaphase spread" was originally used to detect chromosomal abnormalities in cells.
Golgi Method
- (Golgi stain) A selective silver stain technique developed by Camillo Golgi (1843–1926) in 1873.
- This historic technique allowed Santiago Ramón y Cajal (1852–1934) to interpret the structure of the central nervous system.
- There are also a range of other silver staining techniques (see silver staining reticular fibres).
- Links: Neural System Development | Cahal
Gram Stain
A bacterial staining procedure using crystal violet and pink safranin counterstain that generally divides bacteria into either gram-positive or gram-negative and useful for considering associated pharmacology. The procedure was named after Hans Christian Gram (1853 - 1938).
Gram-positive bacteria
- Purple crystal violet stain is trapped by layer of peptidoglycan.
- peptidoglycan forms outer layer of the cell.
Gram-negative bacteria
- Outer membrane prevents stain from reaching peptidoglycan layer in the periplasm.
- outer membrane is composed of four major components: lipopolysaccharide, phospholipids, beta-barrel proteins, and lipoproteins.
- outer membrane then permeabilized.
- Pink safranin counterstain is trapped by peptidoglycan layer.
- Links: Histology Stains | Abnormal Development - Bacterial Infection | Medical Microbiology - Gram stain procedure
Haematoxylin and Eosin
One of the most common staining techniques in pathology and histology.
- Acronym "H and E" stain. (H&E, HE)
- UK - Haematoxylin, USA - Hematoxylin
(Stain - Haematoxylin Eosin) Note - if you have clicked the stain name link where it appears on a page or image on this site it will bring you here, use your web browser back button to return to your original page.
Haematoxylin
- UK - Haematoxylin, USA - Hematoxylin
- Stains nuclei blue to dark-blue.
- Stains the matrix of hyaline cartilage, myxomatous, and mucoid material pale blue.
- Stains myelin weakly but is not noticeable if combined with eosin stain.
Eosin
- Stains cytoplasm pink to red; red blood cells are also bright red.
- Common counterstain to hematoxylin.
- Stain intensity varies with the formula as well as the fixative.
- Eosin - (Greek, eos = dawn, rose-coloured) an acidic dye staining the basic cytoplasmic proteins pink.
- Eosinophil - (Greek, + philein = to love) a type of blood cell with distinct cytoplasmic granules which stain pink with eosin.
- Eosinophilic - having an affinity for eosin dye.
Leishman
- Common blood smear stain.
- Identifies different blood cells.
- identifies infections such as malaria parasites and trypanosomas.
- methylene blue and eosin.
- Links: Practical Lymphoid Tissues
Luxol Fast Blue
- Myelin and phospholipids blue to green.
- copper phthalocyanine dyes.
- Counterstain with Cresyl violet (Nissl stain) to show neuron.
- used as solutions in alcohol or other moderately polar liquids.
- Links: Practical - Neural Histology
Masson’s Trichrome Stain
- Stains nuclei deep blue, skeletal and smooth muscles red, collagen and mucin blue.
- Stains brain and spinal cord parenchymal tissue dusky pink to red.
- Used to evaluate fibrosis
- Striations in skeletal muscles also shows up much better in Masson’s trichrome than in hematoxylin and eosin stain.
- Although called a trichrome, four dyes (hematoxylin, Biebrich scarlet, acid fuchsin, and analine blue) are utilized.
Methenamine Silver
(Jone's Methenamine Silver)
- Stains the basement membrane of the glomerulus in the kidney.
- A routine stain on kidney biopsies.
- Periodic acid oxidizes the carbohydrate components of the basement membrane which produce aldehydes.
- Released aldehydes reduce the silver to a visible metallic silver (black).
- Links: Renal System Histology
Osmium Tetroxide
- has a great affinity for lipids and is chemically bound to the fat and as such acts as a fixative.
- using osmium as a 'stain' makes use of the fact that osmium oxidises tissue fats and forms a black substance which is easily seen in the light microscope.
- adipose tissue, adipose cells, myelin sheaths.
- (Stain - Osmium) Note - if you have clicked the stain name link where it appears on a page or image on this site it will bring you here, use your web browser back button to return to your original page.
Papanicolaou stain
(Papanicolaou's stain, Pap stain) This histology technique was originally described in a publication by George Nikolas Papanicolaou in 1942.[1] A multichromatic (five dyes) staining histological technique has been used to stain many different human bodily fluids (CSF, semen, aspirations), used mainly in the "pap smear" histology. The technique has also been modified several ways (Bismarck brown Y deleted) from the original published technique.
- Haematoxylin - nuclear stain is used to stain cell nuclei.
- Orange G - stains keratin.
- Eosin Y - stains superficial epithelial squamous cells, nucleoli, cilia, and red blood cells.
- Light Green SF yellowish - stains the cytoplasm of non-keratinized squamous cells.
- Bismarck brown Y - stains nothing specifically, often omitted.
- (Stain - Papanicolaou)
Note - if you have clicked the stain name link where it appears on a page or image on this site it will bring you here, use your web browser back button to return to your original page.
Vaginal Smears
- Links: Menstrual Cycle - Histology
Periodic acid-Schiff
(PAS)
- Stains glycogen, mucin, fungus, basement membrane and other substances.
- Stain used to detect fungal organisms and cytoplasmic accumulation of glycogen.
- Stains lysosomes granules red-purple, can be used in recognition of macrophages.
Periodic acid-Schiff / Orange G
- Basophil cells - magenta
- Acidophil cells - yellow
- Red blood cells - yellow
- Nuclei - blue/black
- Chromophobes - pale blue/grey
- Links: Pituitary Histology
Phloxine B
(Phloxine)
- derivatives of eosin
- used in the hematoxylin phloxine saffron (HPS) stain
- stains paneth cell granules in Lendrum's phloxine-tartrazine method
- used to demonstrate alcoholic hyaline
- Formula - C20H2Br4Cl4Na2
PhosphoTungstic Acid Hematoxylin (PTAH)
- Stains nucleus and cytoplasm detail and connective tissue fibers.
- Stains collagen pink, fibrin blue, and striated muscle blue.
- Historic stain used to show CNS reactive astrocytes now used immunochemistry for glial fibrillary acidic protein (GFAP).
Schmorl's Stain
- Stains canaliculi and lamellae in compact bone sections.
- Stain has 2 colouring agents, ammoniacal thionin and aqueous saturated picric acid.
- thionin precipitates within the lacunae and canaliculi (dark brown)
- picric acid forms picrates in the bone matrix (brownish-yellow)
- Named after Christian Georg Schmorl (1861 - 1932) a German pathologist.
Tartrazine
- can be combined with Phloxine and Haem
- tartrazine acts as a yellow counterstain for tissues stained red with phloxine
- pancreatic islets - display pancreatic beta cells
Toluidine Blue
- Stains nucleus blue and cytoplasm light blue.
- A synthetic dye in the thiazins family.
Verhoeff-Van Gieson
- Verhoeff-Van Gieson or elastic-Van Gieson (EVG) stain.
- This is a combination of Verhoeff’s elastic stain which is a hematoxylin stain containing ferric chloride and Wright’s iodine solution and Van Gieson stain which contains acid fuchsin, picric acid, and hematoxylin.
- Stains elastic fibers blue-black to black, collagen pale red, other tissue elements yellow, and nuclei blue to black.
- Named after Ira Thompson Van Gieson (1866 - 1913) an American neurologist and neuropathologist.
- Links: Colon Histology
Whipf's Polychrome
A connective tissue staining technique.
References
- Bancroft JD and Stevens A. Theory and practice of histological techniques. 3rd ed. Churchill Livingstone, 1990.
- Freeman B, Glossary of histological and micro-anatomical terms Department of Anatomy, School of Medical Sciences, UNSW, revised 2000.
External Links
External Links Notice - The dynamic nature of the internet may mean that some of these listed links may no longer function. If the link no longer works search the web with the link text or name. Links to any external commercial sites are provided for information purposes only and should never be considered an endorsement. UNSW Embryology is provided as an educational resource with no clinical information or commercial affiliation.
- Mayo Medical Labs Mallory's Phosphotungstic Acid Hematoxylin (PTAH) Stain
- Sociedad Argentina de Citología Papanicolaou staining protocol
Glossary Links
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Cite this page: Hill, M.A. (2024, June 17) Embryology Histology Stains. Retrieved from https://embryology.med.unsw.edu.au/embryology/index.php/Histology_Stains
- © Dr Mark Hill 2024, UNSW Embryology ISBN: 978 0 7334 2609 4 - UNSW CRICOS Provider Code No. 00098G